Molecular marker GI1354 closely linked with pepper CMV resistance gene and acquisition and application

A molecular marker and resistance gene technology, applied in the field of genetic engineering, can solve the problem that there is no single recessive gene molecular marker for pepper CMV resistance, and achieve the effect of convenient method and cost saving.

Inactive Publication Date: 2016-03-02
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far, there is no relevant research report on the molecular markers closely linked to the pepper CMV resistance single recessive gene

Method used

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  • Molecular marker GI1354 closely linked with pepper CMV resistance gene and acquisition and application
  • Molecular marker GI1354 closely linked with pepper CMV resistance gene and acquisition and application
  • Molecular marker GI1354 closely linked with pepper CMV resistance gene and acquisition and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] This example is the breeding method and identification method of pepper material Q132 controlled by a single recessive gene for CMV resistance. On the basis of obtaining the pepper material Q132 whose CMV resistance is controlled by a single recessive gene, it is possible to obtain and develop molecular markers closely linked to the pepper CMV resistance gene.

[0042] Wherein, the above-mentioned breeding method comprises the following steps, such as figure 1 shown.

[0043] (1) Selection of hybrid materials: Select four high-quality pepper hybrids introduced from home and abroad: "37-74", "Xiyangyang", "Pavor", and "Sao Paulo".

[0044] Among them, "37-74" was purchased from Rexwang (China) Seed Co., Ltd., "Xiyangyang" was purchased from Changle Xinsheng Seed Industry Co., Ltd., and "Pavol" was purchased from Japan Sakata Seedling (Suzhou) Co., Ltd. "St. Paul" was purchased from Nunhem (Beijing) Seed Co., Ltd.

[0045] (2) Hybridization: pairwise hybridization was ...

Embodiment 2

[0110] On the basis of the capsicum material Q132 whose CMV resistance is controlled by a single recessive gene has been obtained in Example 1, this embodiment is a method for obtaining the molecular marker genSSR3000 closely linked to the capsicum CMV resistance gene, including the following steps.

[0111] (1) Linkage marker analysis:

[0112] ① Extraction of genomic DNA:

[0113] Using Tiangen Plant Genomic DNA Extraction Kit (DP305-2) to extract parents, F 2 , B 1 For the genomic DNA of each individual plant in the population, refer to the operating instructions of the kit for details on the extraction method, and obtain the genomic DNA of each plant.

[0114] ②Development of a large number of SSR primers and InDel primers:

[0115] Using the genome sequence (Version1.55) (Kimetal.2014) of pepper CM334 as a reference sequence, develop the SSR site, use the Perl program to develop the SSR site in the protein-coding gene region and its upstream and downstream regions of 5...

Embodiment 3

[0155] This example is the application of the above-mentioned molecular marker GI1354 closely linked to the cr gene to identify whether the cr gene is contained in capsicum materials with different genetic backgrounds.

[0156] The above-mentioned molecular marker GI1354 was used as a PCR primer to carry out PCR amplification reaction on the genomic DNA of different pepper materials to obtain the amplified product, and then the amplified product was subjected to non-denaturing polyacrylamide gel electrophoresis, and after silver staining, the banding Type statistical analysis to determine whether pepper material contains cr gene.

[0157] The above-mentioned molecular marker GI1354 closely linked with the cr gene is applied to the method for identifying whether the capsicum material contains the cr gene, including the following steps:

[0158] (1) Genomic DNA extraction of pepper material: Genomic DNA of pepper material was extracted using Tiangen Plant Genomic DNA Extraction ...

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Abstract

The invention belongs to the field of gene engineering, and particularly relates to a molecular marker GI1354 closely linked with a pepper CMV resistance gene and an acquisition method and application. The molecular marker GI1354 has at least one of nucleotide base sequences shown in SEQ ID NO:1 and SEQ ID NO:2 in the sequence list. During detection of the cr gene in the molecular marker application pepper material, the steps of extraction of genome DNA of the pepper material, the PCR amplification reaction, analysis and the like are included. The screened PCR primer is directly used as the molecular marker, whether the cr gene is contained in the pepper material or not can be accurately detected, and the method is convenient to implement, rapid and capable of saving cost.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a molecular marker GI1354 closely linked with a capsicum CMV resistance gene, an acquisition method and application. Background technique [0002] Pepper (Capicumannuum L.) is an important vegetable crop widely cultivated worldwide. Cucumber mosaic virus (CMV) is a devastating disease and one of the main viruses that harm pepper. CMV often causes deformity and shrinkage of pepper leaf shape, and in severe cases, the plant shrinks, and the fruit is small and hard, resulting in a decrease in yield and quality. CMV can seriously affect the yield and quality of pepper. In general years, it can cause a 20%-30% reduction in pepper production, and in severe cases it can reach 50%-60%. The chemical pesticides frequently used in the cultivation and production process to prevent and control pepper CMV disease have affected the edible safety of pepper products, posed a serio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 张晓芬耿三省陈斌
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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